Study On The Chemical Composition And Quality Standard Of Hollyhock Seeds

Althaea rosea(Linn.)Cavan is a biennial erect herb of the Althaea rosea genus Malvaceae.It is mainly grown and distributed in Henan,Hebei,Gansu,Xinjiang,Sichuan and other places.It is a traditional ethnic medicine and is used in the Uygur,Bai,WA,Naxi and so on.The root has the functions of clearing away heat,detoxification,purulent discharge and diuresis;Seeds can make the urinary tract leaching;Flowers can make the stool smooth,detoxify and stagnate;flowers and leaves are used externally to treat boils,sores and burns.The research on the chemical composition of Althaea rosea,and found that the known chemical constituents of Althaea rosea mainly included flavonoids,phenolic acids,steroids,phenylpropanoids,sugars and other constituents.Modern pharmacological studies have confirmed that Althaea rosea has anti-inflammatory and analgesic,antioxidant,bacteriostatic,antidepressant,protective effect on renal tubular injury,and pharmacological effects such as immune regulation.The purpose of this study was to study the chemical constituents of Althaea rosea seeds,establish the HPLC fingerprinting of Althaea rosea seeds and determine the content of active ingredients in them,which will lay the foundation for the further development of Althaea rosea seeds quality standards and the rational development and utilization.This paper is divided into two parts.Part ?:study on chemical constituents of Althaea rosea seedsPolyamide column chromatography,Reverse silica gel column chromatography,Silica gel column chromatography,HPLC separation methods were used,along with the MS,LC-MS,NMR and other identification methods were used to study Althaea rosea seeds.11 compounds were identified from Althaea rosea seeds,containing two flavonoids compounds,eight phenolic compounds,one alkaloid compound.Two flavonoids compounds were Vicenin-1(1),Schaftoside(2).Eight phenolic compounds were p-hydroxy-phenylpropionic acid(3),dihydroferulic acid(4),p-hydroxyphenethyl(5),3,4-dimethoxy-benzoic acid(6),Syringic acid(7),N-ethyl-4-hydroxy-3-methoxybenzamide(8),4-Hydroxybenzoic acid(9),3-Hydroxy-4-methoxybenzoic acid(10).One alkaloid compound was pyridinecarboxylic acid(11).11 compounds were isolated from this plant for the first time.Part ?:Establishment of HPLC Fingerprinting of Althaea rosea seeds and determination of active ingredientsFirst,reversed-phase high performance liquid chromatography was used to establish corresponding HPLC fingerprinting of Althaea rosea seeds from different regions.The chromatographic conditions were Agilent Pursuit 5 C₁₈(4.6×250 mm,5?m)chromatographic column.The mobile phase A is methanol,and the volume fraction of phase B is 0.2%phosphoric acid aqueous solution.Gradient elution(0?30min,10%?45%A,30?32min,45%?10%A,32?40min,10%?10%A).The flow rate was 1.0m L/min,the detection wavelength was 254nm,and the column temperature was 30?.The reference was schaftoside.Results there was a small difference in fingerprint similarity between 15 batches of Althaea rosea seeds from different areas.And the comparison fingerprint of Althaea rosea seeds was generated by using the similarity evaluation software of traditional Chinese medicine chromatographic fingerprint.There were 10 chromatographic peaks,each peak of which was well separated and met the requirements of fingerprint detection,which could be used as the main basis for the identification of Althaea rosea seeds.The established Althaea rosea seeds fingerprint could be used for the identification of varieties and quality evaluation of Althaea rosea seeds.Secondly,the contents of two components in Althaea rosea seeds were determined simultaneously by reversed-phase high performance liquid chromatography.Using Vicenin-1 and schaftoside as control substances(both compounds were extracted from Althaea Rosea Seeds in this experiment,and their purity was greater than 98.0%by high performance liquid chromatography area normalization method);The chromatographic conditions were Agilent Pursuit 5 C₁₈(4.6×250 mm,5?m)chromatographic column.The mobile is methanol(phase A)-0.1%formic acid solution(phase B),Gradient elution(0?8 min,30%?32%A,8?20min,32%?34%A,20?21 min,34%?30%A,21?30 min,30%?30%A),flow rate of1.0 m L·min.The detection Wavelength was set at 272 nm and the column temperature was 30?.The method showed a good linear relationship within the range of0.0195?0.234?g(r=0.9998)for vicenin-1and 0.0653?1.632?g(r=0.9998)for schaftoside;The RSD of the repeatability of vicenin-1 and schaftoside(n=6)were1.79%and 1.92%;the precision(n=6)RSD is 0.27%and 0.19%;the stability(n=6)RSD is 1.00%and 0.60%;the average recovery rate(n=6)is 100.51%(RSD=1.03%),100.55%(RSD=0.55%);The content of vicenin-1 ranged from 0.072%to0.111%,and that of schaftoside ranged from 0.044%to 0.068%.The experimental results show that the method is simple,rapid,accurate and repeatable,It can be applied to the determination of Althaea rosea seeds content,which provides a reference for the more effective control of Althaea rosea seeds quality and the rational development and utilization of Althaea rosea seeds.